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苜蓿中华根瘤菌中影响非氧化戊糖磷酸途径的突变的表征

Characterization of Mutations That Affect the Nonoxidative Pentose Phosphate Pathway in Sinorhizobium meliloti.

作者信息

Hawkins Justin P, Ordonez Patricia A, Oresnik Ivan J

机构信息

Department of Microbiology, University of Manitoba, Winnipeg, Manitoba, Canada.

Department of Microbiology, University of Manitoba, Winnipeg, Manitoba, Canada

出版信息

J Bacteriol. 2017 Dec 20;200(2). doi: 10.1128/JB.00436-17. Print 2018 Jan 15.

Abstract

is a Gram-negative alphaproteobacterium that can enter into a symbiotic relationship with and Previous work determined that a mutation in the gene, which encodes a putative transketolase, could prevent medium acidification associated with a mutant strain unable to metabolize galactose. Since the pentose phosphate pathway in is not well studied, strains carrying mutations in either and , which encodes a putative transaldolase, were characterized. Carbon metabolism phenotypes revealed that both mutants were impaired in growth on erythritol and ribose. This phenotype was more pronounced for the mutant strain, which also displayed auxotrophy for aromatic amino acids. Changes in pentose phosphate pathway metabolite concentrations were also consistent with a mutation in either or The concentrations of metabolites in central carbon metabolism were also found to shift dramatically in strains carrying a mutation. While the concentrations of proteins involved in central carbon metabolism did not change significantly under any conditions, the levels of those associated with iron acquisition increased in the wild-type strain with erythritol induction. These proteins were not detected in either mutant, resulting in less observable rhizobactin production in the mutant. While both mutants were impaired in succinoglycan synthesis, only the mutant strain was unable to establish symbiosis with alfalfa. These results suggest that and play central roles in regulating the carbon flow necessary for carbon metabolism and the establishment of symbiosis. is a model organism for the study of plant-microbe interactions and metabolism, especially because it effects nitrogen fixation. The ability to derive the energy necessary for nitrogen fixation is dependent on an organism's ability to metabolize carbon efficiently. The pentose phosphate pathway is central in the interconversion of hexoses and pentoses. This study characterizes the key enzymes of the nonoxidative branch of the pentose phosphate pathway by using defined genetic mutations and shows the effects the mutations have on the metabolite profile and on physiological processes such as the biosynthesis of exopolysaccharide, as well as the ability to regulate iron acquisition.

摘要

是一种革兰氏阴性α-变形菌,能够与[具体对象1]和[具体对象2]建立共生关系。先前的研究确定,编码假定转酮醇酶的[基因名称]基因发生突变,可能会阻止与无法代谢半乳糖的突变菌株相关的培养基酸化。由于[研究对象]中的磷酸戊糖途径尚未得到充分研究,因此对携带[基因名称1]和[基因名称2]突变的菌株进行了表征,其中[基因名称2]编码假定转醛醇酶。碳代谢表型显示,这两种突变体在赤藓糖醇和核糖上的生长均受到损害。这种表型在[基因名称1]突变菌株中更为明显,该菌株还表现出对芳香族氨基酸的营养缺陷。磷酸戊糖途径代谢物浓度的变化也与[基因名称1]或[基因名称2]的突变一致。在携带[基因名称]突变的菌株中,也发现中心碳代谢中的代谢物浓度发生了显著变化。虽然参与中心碳代谢的蛋白质浓度在任何条件下都没有显著变化,但在野生型菌株中,随着赤藓糖醇诱导,与铁获取相关的蛋白质水平增加。在这两种突变体中均未检测到这些蛋白质,导致[基因名称1]突变体中根瘤菌素的产生较少。虽然这两种突变体在琥珀聚糖合成方面均受到损害,但只有[基因名称1]突变菌株无法与苜蓿建立共生关系。这些结果表明,[基因名称1]和[基因名称2]在调节碳代谢和共生关系建立所需的碳流中起核心作用。[研究对象]是研究植物-微生物相互作用和代谢的模式生物,特别是因为它影响固氮作用。获得固氮所需能量的能力取决于生物体有效代谢碳的能力。磷酸戊糖途径在己糖和戊糖的相互转化中起核心作用。本研究通过使用特定的基因突变来表征磷酸戊糖途径非氧化分支的关键酶,并展示了这些突变对代谢物谱以及对外多糖生物合成等生理过程的影响,以及调节铁获取的能力。

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